Advanced Microscopic Techniques and Nanotechnology

A.Y. 2025/2026
6
Max ECTS
51
Overall hours
SSD
FIS/07
Language
English
Learning objectives
The course is meant to be an introduction to biotechnologically relevant techniques that requires a particular knowledge of their physical, computational and technological basis.

The course aims at giving describing the principles and the potentials of a selection of advanced optical microscopy techniques and of nanotechnologies. The course also aims at providing a basic understanding and training of quantitative image analysis.

The topics included in the course can be divided into three main sections.
Expected learning outcomes
In this class the student: (i) learns about the physical quantities of importance in the field of Biotechnologies, with a specific emphasis on thermodynamic quantities, and about their units of measurements; (ii) learns to use such quantities in quantitative problems; (iii) learns about the physical and technological foundations of optical microscopy and fluorescence.
Single course

This course cannot be attended as a single course. Please check our list of single courses to find the ones available for enrolment.

Course syllabus and organization

Single session

Lesson period
First trimester
Course syllabus
ADVANCED OPTICAL MICROSCOPY

Introduction on the resolution of optical microscopes
Effects of the finite resolution on images
Introduction to confocal microscopy
Optical processes and techniques that can overcome the resolution limit, such as: non linear microscopy, STED,TIRF,PALM, SNOM.
Introduction to time resolved fluorescence and FLIM microscopy

NANOTCHNOLOGY

Aims and uses of nanoparticles in biomedicine.
Nanoparticles: quantum dots, nanomag, metallic nanoparticles, polymer particles, liposomes.
General concepts, stability, bioconiugation, cell internalization.
Optical tweezers for micro-manipulation.
Atomic force microscopy.
Later-free optical biosensors. Introduction to SPR-based techniques.
Micromechanical devices.

QUANTITATIVE ANALYSIS OF IMAGES

Introduction to the fundamentals of computer graphics aimed to the understanding and elaboroation of the informations contained in images.
Colorimetry: color spectrum, Gamut, chromatic coordinates, gamma value of displays, RGB, CMYK.
Digital image types: (es. BMP,TIF,GIF,JPG)
Lossy and lossless compression
ImageJ interface introduction, Image visualization (Look Up Tables, Brightness and Contrast), Pixel Statistics.
Processing examples (FFT and filtering)
An example of a quantitative analysis: electrophoretic gel
Object recognition I : image segmentation and particle analysis
Multidimensional images from color channels to image stacks
Video processing
Best Fitting procedures. Fitting a model into quantitative data extracted from a digital image. Extraction of experimental parameters and confidence levels.
Object Recognition II: Training a convolutional Netural Networks for the automatic recognition of objects in image. Image annotation.
Prerequisites for admission
Students are required to have some knowledge of: basics of optics (basic properties of light: speed, frequency, wavelength, refraction), basic properties of lenses (focal length, image formation), basic structure of the optical microscopes (objective, eyepiece, magnification, upright or inverted structure, illumination). Those who do not have these elements of knowledge are required of an effort of personal study. During the first hour of the class, the required elements of knowledge will be listed, together with web sites that can be used for the study material, in case no other reference book is available to the students.
Teaching methods
The course comprised three main topics: microscopy, nanotechnology, and digital images. The first two are given through lessons, while the third is developed in a combination of lessons and hands-on experiences. Students will be asked to analyze digital images made available by the teachers through the Ariel platform. Hands-on exercises will be carried on by single students or by pairs of students, as they prefer. For the exercises it will be possible to use the computers own by the students. This will be discussed and decided by the students during the first lesson.
The teaching material includes: pdf files with the slides used during the lessons and articles. The articles are either review papers or applications of the various technologies introduced in the class. Within slides students will find comments - not used during the lessons - that summarize the contents, aimed at helping studying.
Teaching Resources
On the course website you will find the files of the slides used in the lectures and a selection of review articles on topics drawn from important international magazines. The part of the hands-on practice using the ImageJ software that is available online without the need for licenses.
Assessment methods and Criteria
The exam will be in written form and composed of six open questions involving both the understanding of the basics of the technologies and exemplification of their use.
The questions will be not too dissimilar (but not identical) to those in a list made available to the students from the myAriel website.
The total duration of the exam will be of about 1.5 hours.
FIS/07 - APPLIED PHYSICS - University credits: 6
Practicals: 16 hours
Lessons: 35 hours